Team:Warsaw/Calendar-Main/21 August 2008

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Revision as of 17:14, 20 October 2008


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OmpA variant ("hunter")	"Prey" variant	Growth with prey	Growth without prey
OmpA-alpha	His+Z+omega	+	+
OmpA-alpha	His+Z+alfa	+	+
OmpA-A-alpha	His+Z+omega	+	-
OmpA-Z-omega	His+A+alpha	+	-
OmpA-omega-A-alpha		+	+
Ompa-A-omega	His+Z+alpha	+	-
Ompa-A-omega	His+Z+omega	-	-
OmpA-omega	His+Z+alpha	+	-
Ompa-Z-alpha	His+Z+omega	+	-
OmpA_Adelta_alpha	His+Z+omega	+	-
OmpA_Adelta_omega	His+Z+alpha	+	-
OmpA_omega_Adelta	His+Z+alpha	+	-

Cloning of protein A DNA to GeneArt plasmid

Antoni

Colony PCR on colonies from plates with transformations pGeneart+A.
Primers used: AP+NotI and AL+SacI.
Gel electrophoresis.
Confirmed transformant colonies inoculated to liquid LB with ampicillin.
Inoculation to liquid LB with ampicillin: pET15b+OmpA-alpha.

Cloning of alpha to pACYC177+OmpA_A_omega and pACYC177+OmpA_Z_omega

Michał K.

Isolation of plasmids from cultures inocluated on previous day.
Control digest of isolated plasmids with FastBamHI and FastAcc65I (Fast Digest buffer).
Gel electrophoresis - no proper clones founded.

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-OmpA_A_alpha, OmpA_Adelta_alpha, OmpA_Adelta_omega, OmpA_A_omega, OmpA_omega_Adelta, OmpA_omeg
 <table id="result">
 <tr><th>OmpA variant ("hunter")</th><th>"Prey" variant</th><th>Growth with prey</th><th>Growth without prey</th></tr>
-<tr><th>OmpA-alpha</th><td>His+Z+omega</td><td>no</td><td>no</td></tr>
+<tr><th>OmpA-alpha</th><td>His+Z+omega</td><td>+</td><td>+</td></tr>
-<tr><th>OmpA-alpha</th><td>His+Z+alfa</td><td>no</td><td>no</td></tr>
+<tr><th>OmpA-alpha</th><td>His+Z+alfa</td><td>+</td><td>+</td></tr>
-<tr><th>OmpA-A-alpha</th><td>His+Z+omega</td><td>yes</td><td>no</td></tr>
+<tr><th>OmpA-A-alpha</th><td>His+Z+omega</td><td>+</td><td>-</td></tr>
-<tr><th>OmpA-Z-omega</th><td>His+A+alpha</td><td>yes</td><td>no</td></tr>
+<tr><th>OmpA-Z-omega</th><td>His+A+alpha</td><td>+</td><td>-</td></tr>
-<tr><th>OmpA-omega-A-alpha</th><td>His+Z+omega</td><td>yes</td><td>no</td></tr>
+<tr><th>OmpA-omega-A-alpha</th><td></td><td>+</td><td>+</td></tr>
-<tr><th>Ompa-A-omega</th><td>His+Z+alpha</td><td>yes</td><td>no</td></tr>
+<tr><th>Ompa-A-omega</th><td>His+Z+alpha</td><td>+</td><td>-</td></tr>
-<tr><th>Ompa-A-omega</th><td>His+Z+alpha</td><td>yes</td><td>no</td></tr>
+<tr><th>Ompa-A-omega</th><td>His+Z+omega</td><td>-</td><td>-</td></tr>
-<tr><th>OmpA-omega</th><td>His+A+alpha</td><td>no</th><td>no</td></tr>
+<tr><th>OmpA-omega</th><td>His+Z+alpha</td><td>+</th><td>-</td></tr>
-<tr><th>Ompa-Z-alpha</th><td>His+A+omega</td><td>yes</td><td>no</td></tr>
+<tr><th>Ompa-Z-alpha</th><td>His+Z+omega</td><td>+</td><td>-</td></tr>
-<tr><th>Ompa-Z-alpha</th><td>His+A+omega</td><td>yes</td><td>no</td></tr>
+<tr><th>OmpA_Adelta_alpha</th><td>His+Z+omega</td><td>+</td><td>-</td></tr>
+<tr><th>OmpA_Adelta_omega</th><td>His+Z+alpha</td><td>+</td><td>-</td></tr>
+<tr><th>OmpA_omega_Adelta</th><td>His+Z+alpha</td><td>+</td><td>-</td></tr>
 </table>